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An advanced molecular strategy to identify bacterial communities on art objects
C Schabereiter-Gurtner1, G Piñar, W Lubitz
1Institute of Microbiology and Genetics, University of Vienna, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria. gurtifox@gem.univie.ac.at
Journal of Microbiological Methods
|April 20, 2001
Summary
This study presents a new DNA extraction protocol for analyzing bacterial communities on art objects using molecular methods. The protocol enables more reliable identification of microorganisms compared to traditional techniques.
Area of Science:
- Microbiology
- Molecular Biology
- Conservation Science
Background:
- Conventional cultivation methods underestimate bacterial diversity on art objects.
- Molecular techniques like PCR amplification of 16S rRNA genes offer deeper insights.
- DNA extraction from small art object samples with PCR inhibitors is challenging.
Purpose of the Study:
- To develop and validate a novel DNA extraction protocol for microbial analysis of art objects.
- To enable reliable phylogenetic identification of microorganisms from challenging samples.
- To overcome limitations of conventional cultivation and direct sequencing of DGGE bands.
Main Methods:
- Developed a DNA extraction protocol for lime wall paintings and loamy soil.
- Utilized PCR amplification of 16S rRNA gene fragments.
- Employed Denaturing Gradient Gel Electrophoresis (DGGE) for analysis.
- Constructed and screened clone libraries for phylogenetic analysis.
Main Results:
- Successfully extracted PCR-amplifiable DNA from art object samples.
- Identified complex bacterial communities beyond cultivation capabilities.
- Demonstrated superior phylogenetic resolution using clone libraries compared to direct DGGE band sequencing.
Conclusions:
- The new DNA extraction protocol effectively supports molecular investigation of microbial communities on art objects.
- Clone libraries provide more reliable microorganism identification than direct DGGE sequencing.
- Culture-independent molecular methods significantly advance the study of microbial diversity in cultural heritage.